Apoptotic role of microsomal glucose-6-phosphate in granulocytes  Page description

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Details of project

 
Identifier
46740
Type F
Principal investigator Kardon, Tamás Zoltán
Title in Hungarian Mikroszómális glukóz-6-foszfát szerepe granulocita apoptózisában
Title in English Apoptotic role of microsomal glucose-6-phosphate in granulocytes
Panel Cellular and Developmental Biology
Department or equivalent Dept. of Medical Chemistry, Molecular Biology and Pathobiochemistry (Semmelweis University)
Participants Csala, Miklós
Nagy, Gábor
Starting date 2004-01-01
Closing date 2008-12-31
Funding (in million HUF) 5.199
FTE (full time equivalent) 0.00
state closed project





 

Final report

 
Results in Hungarian
Az endoplazmás retikulum glukóz-6-foszfát transzporterére specifikus poliklonális antitestet termeltettünk, amely patkány és humán glukóz-6-foszfát transzportert ismer fel. Western blottal és immunhisztomkémiai módszerekkel közvetlenül bizonyítottuk a transzporter jelenlétét humán májból, patkány májból és veséből származó mikroszómákban; humán granulocitákban, patkány agyban és harántcsíkolt izomszövetben. Kimutattuk, hogy humán granulociták és differenciálódott HL60-as sejtekben megtalálható a hexóz-6-foszfát dehidrogenáz, amelynek a működése függ a glukóz-6-foszfáttól, aktív centruma az endoplazmás retikulum lumenében található és NADPH-t generál. A keletkezett 6-foszfoglukonátot a granulocita mikroszómák nem metabolizálják tovább. mRNS és fehérje szinten kimutattuk 1-es típusú 11-béta-hidroxiszteriod dehidrogenáz jelenlétét granulociták mikroszómáiban. Az enzim granulocitákban is oxidoreduktázként működik, működéséhez intraluminális NADPH(NADP)-t használ fel. Az enzim NADPH-t generáló kortizol dehidrogenáz aktivitásával kivédtük a glukóz-6-foszfát transzport gátlásával létrehozott apoptózist. Eredményeink szerint a májhoz hasonlóan humán granulocitákban is megtalálható a glukóz-6-foszfát - hexóz-6-foszfát és 11-béta-hidroxiszeriod dehidrogenáz 1 funkcionális egysége.
Results in English
We employed antibodies raised against the liver glucose-6-phoshphate transporter protein. With different immunodetection methods, we have shown that the protein is expressed in endoplasmic reticulum membranes derived from human and rat liver and rat kidney. The expression was also present in the following tissues: human granulocytes, rat brain and skeletal muscle. We have demonstrated the expression of hexose-6-phosphate dehydrogenase in human neutrophils and mature HL60-cells. Its catalytic subunit is intraluminal and activity of the enzyme was shown to be dependent on glucose-6-phosphate and produces NADPH. 6-phosphogluconate formed in this reaction was not metabolized further. The expression and activity of 11-beta-hydroxysteroid dehydrogenase type 1, another NADP(H)-dependent microsomal enzyme were also detected in human neutrophils. The NADPH-generating cortisol dehydrogenase activity of the enzyme prevented neutrophil apoptosis provoked by the inhibition of the glucose-6-phosphate transporter. The results show that granulocytes are equipped with a functional glucose-6-phosphate- hexose-6-phosphate dehydrogenase-11beta-hydroxysteroid dehydrogenase type 1 system.
Full text http://real.mtak.hu/1559/
Decision
Yes





 

List of publications

 
Senesi S; Marcolongo P; Kardon T; Bucci G; Sukhodub AL; Burchell A; Benedetti A; Fulceri R: Gene Products of the Glucose-6-phosphate Transporter Gene, In: Benedetti A, Bánhegyi G, Burchell A (ed.) Endoplasmic Reticulum: A Metabolic Compartment, Amsterdam: IOS Press, 2005. pp. 21-29, 2005
Senesi S; Marcolongo P; Kardon T; Bucci G; Sukhodub A; Burchell A; Benedetti A; Fulceri R: Immunodetection of the expression of microsomal proteins encoded by the glucose 6-phosphate transporter gene, BIOCHEMICAL JOURNAL 389: 57-62, 2005
Legeza B; Banhegyi G; Mandl J; Benedetti A; Kardon T;: The presence of 11 beta-hydroxysteroid dehydrogenase type I in human granulocytes, FEBS Journal 272 (s1), 490; L1-022P, 2005
Kardon T; Leuzzi R; Marcolongo P; Senesi S; Bánhegyi G; Burger HJ; Benedetti A; Fulceri R;: What is the Role for the Glucose-6-Phosphate Transporter in Leukocytes?, In: Benedetti A, Bánhegyi G, Burchell A (ed.) Endoplasmic Reticulum: A Metabolic Compartment, Amsterdam: IOS Press, 2005. pp. 30-35, 2005
Czegle I, Piccirella S, Senesi S, Csala M, Mandl J, Banhegyi G, Fulceri R, Benedetti A.: Cooperativity between 11beta-hydroxysteroid dehydrogenase type 1 and hexose-6-phosphate dehydrogenase is based on a common pyridine nucleotide pool in the lumen of the endoplasmic reticulum, Mol Cell Endocrinol. 248;24-25., 2006
Piccirella S, Czegle I, Lizak B, Margittai E, Senesi S, Papp E, Csala M, Fulceri R, Csermely P, Mandl J, Benedetti A, Banhegyi G.: Uncoupled redox systems in the lumen of the endoplasmic reticulum. Pyridine nucleotides stay reduced in an oxidative environment., J Biol Chem 281;4671-4677, 2006
Legeza B, Senesi S, Marcolongo P, Bánhegyi G, Fulceri R, Benedetti A, Kardon T: Generating NADPH in the Endoplasmic Reticulum of Granulocytes: A Possible Antiapoptotic Mechanism, 31. FEBS Congress Istanbul, Abstract book, 2006
Lizák B; Czegle I; Csala M; Benedetti A; Mandl J; Bánhegyi G.: Translocon pores in the endoplasmic reticulum are permeable to small anions., Am J Physiol Cell Physiol. 291:C511-7., 2006
Bánhegyi G; Benedetti A, Csala M; Mandl J;: Stress on redox, FEBS Lett. 581:3634-40., 2007
Lizák B; Csala M; Benedetti A; Bánhegyi G.: The translocon and the non-specific transport of small molecules in the endoplasmic reticulum, Molecular Membrane Biology, 2008




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